title: Crystallization and preliminary X-ray crystallographic study of a methyltransferase involved in 2-methylisoborneol biosynthesis in Streptomyces lasaliensis.
abstract: The biosynthetic pathway of the off-flavour terpenoid alcohol 2-methylisoborneol (2-MIB) requires geranyl pyrophosphate methyltransferase (GPPMT) to methylate GPP before the cyclization reaction. GPPMT is the first example of an S-adenosyl-L-methionine-dependent methyltransferase that acts on general intermediates such as geranyl pyrophosphate and farnesyl pyrophosphate in isoprenoid biosynthetic pathways. In this study, recombinant GPPMT was overproduced, purified and crystallized in the absence and presence of cofactor, cofactor analogue and substrate. Well diffracting crystals of apo GPPMT containing one molecule in the asymmetric unit were obtained and the structure of this form was solved by the molecular-replacement method. Two crystal forms of the tertiary complex with GPP and sinefungin were also obtained. Structure analysis of these crystals is currently under way in order to understand the enzyme reaction mechanism.
authors.affiliation: Graduate School of Life Sciences, Hokkaido University, Sapporo 060-0810, Japan.
authors.forename: Orapin
authors.initials: O
authors.lastname: Ariyawutthiphan
authors.forename: Toyoyuki
authors.initials: T
authors.lastname: Ose
authors.forename: Muneya
authors.initials: M
authors.lastname: Tsuda
authors.forename: YongGui
authors.initials: Y
authors.lastname: Gao
authors.forename: Min
authors.initials: M
authors.lastname: Yao
authors.forename: Atsushi
authors.initials: A
authors.lastname: Minami
authors.forename: Hideaki
authors.initials: H
authors.lastname: Oikawa
authors.forename: Isao
authors.initials: I
authors.lastname: Tanaka
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D001892:Camphanes / Q000737:chemistry / Q000378:metabolism*; D003460:Crystallization; D018360:Crystallography, X-Ray; D008780:Methyltransferases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D015394:Molecular Structure; D011106:Polyisoprenyl Phosphates / Q000737:chemistry / Q000378:metabolism; D013302:Streptomyces / Q000737:chemistry / Q000201:enzymology*
chemical_list: D001426:Bacterial Proteins; D001892:Camphanes; D011106:Polyisoprenyl Phosphates; C005536:2-methylisoborneol; C015234:geranyl pyrophosphate; D008780:Methyltransferases